目的:探讨氟、砷及氟砷联合染毒对小鼠成骨细胞MC3T3-E1与小鼠单核巨噬细胞RAW264.7共培养体系中肿瘤坏死因子受体相关因子6（TRAF-6）/核因子κB1（NF-κB1）信号通路中相关蛋白表达的影响。方法:MC3T3-E1细胞经成骨诱导剂诱导后与RAW264.7细胞建立共培养体系，体外培养7 d，采用析因设计，分别用不同剂量的氟化钠（0.0、0.1、0.4、1.6 mmol/L NaF，F）、亚砷酸钠（0.0、0.5、2.5、12.5 μmol/L NaAsOn 2，As）以及不同剂量的氟砷联合培养基培养24 h。采用蛋白免疫印迹法（Western blot）检测核因子κB受体活化因子（RANK）、TRAF-6、NF-κB1、T细胞活化因子（NFATc1）、抗酒石酸酸性磷酸酶（TRAP）的蛋白表达水平。n 结果:氟单独作用时，与对照组（Fn 0.0Asn 0.0，1.00 ± 0.00）比较，各剂量组RANK、NF-κB1、TRAP蛋白表达（1.11 ± 0.04、1.29 ± 0.05、1.38 ± 0.04，1.24 ± 0.04、1.13 ± 0.03、1.34 ± 0.05，1.12 ± 0.03、1.24 ± 0.04、1.61 ± 0.06）增加（n P均< 0.05）；TRAF-6蛋白表达在Fn 0.1和Fn 1.6组（1.23 ± 0.04、1.35 ± 0.03）增加（n P均< 0.05）。砷单独作用时，与对照组（Fn 0.0Asn 0.0）比较，RANK、TRAF-6、NF-κB1蛋白表达在Asn 0.5组增加（n P均< 0.05），RANK和NFATc1蛋白表达在Asn 12.5组减少（n P均< 0.05）。氟砷联合作用时，同一染氟剂量，RANK蛋白表达在Fn 0.1Asn 0.5组，TRAF-6蛋白表达在Fn 0.1Asn 12.5、Fn 0.4Asn 0.5、Fn 0.4Asn 2.5组，NF-κB1蛋白表达在Fn 0.1Asn 0.5、Fn 0.4Asn 2.5、Fn 0.4Asn 12.5组，NFATc1蛋白表达在Fn 0.1Asn 0.5、Fn 0.4Asn 0.5组，TRAP蛋白表达在Fn 0.1Asn 12.5组均高于相应的单独氟染毒组（Fn 0.1、Fn 0.4，n P均< 0.05），但低于氟、砷单独染毒之和。同一染砷剂量，RANK蛋白表达在Fn 0.1Asn 12.5组，TRAF-6蛋白表达在Fn 0.1Asn 12.5、Fn 0.4Asn 2.5组，NF-κB1蛋白表达在Fn 0.1Asn 12.5、Fn 0.4Asn 2.5、Fn 0.4Asn 12.5、Fn 1.6Asn 2.5组，TRAP蛋白表达在Fn 1.6Asn 2.5、Fn 1.6Asn 12.5组均高于相应的单独砷染毒组（Asn 2.5、Asn 12.5，n P均< 0.05），但低于氟、砷单独染毒之和。氟对RANK、TRAF-6、NF-κB1、NFATc1、TRAP蛋白表达均有主效应作用（n F=3.41、341.73、66.01、56.49、147.40，n P均< 0.05）；砷对各蛋白指标也均有主效应作用（n F=686.71、174.96、107.32、235.80、331.37，n P均< 0.05）；氟砷联合作用对各蛋白指标均有交互作用（n F=50.39、234.94、116.72、67.77、36.56，n P均< 0.05）。n 结论:在MC3T3-E1与RAW264.7细胞的共培养体系中，氟可激活TRAF-6介导的NF-κB1信号通路相关蛋白表达，从而促进破骨细胞分化；砷对相关蛋白表达作用不完全一致。氟砷联合染毒对TRAF-6介导的NF-κB1信号通路相关蛋白表达的交互作用主要表现为拮抗作用。“,”Objective:To investigate the effects of combined exposure of fluorine, arsenic, and fluorine-arsenic on the signaling pathway related protein expression of tumor necrosis factor receptor-related factor 6 (TRAF-6)/nuclear factor κB1(NF-κB1) in a co-culture system of mouse osteoblasts MC3T3-E1 and mouse monocyte macrophage RAW264.7.Methods:MC3T3-E1 cells were co-cultured with RAW264.7 cells after induction with osteogenic inducers. The cells were cultured for 7 days n in vitro, and different doses of sodium fluoride (0.0, 0.1, 0.4, 1.6 mmol/L NaF, F), sodium arsenite (0.0, 0.5, 2.5, 12.5 μmol/L NaAsO n 2, As) and different doses of fluorine and arsenic were added to the culture medium and cultured for 24 h using factorial design. The expression levels of nuclear factor κB receptor activating factor (RANK), TRAF-6, NF-κB1, T cell activating factor (NFATc1), and tartrate-resistant acid phosphatase (TRAP) protein were detected by Western blotting.n Results:When fluorine was used alone, compared with the control group (Fn 0.0Asn 0.0, 1.00 ± 0.00), the expressions of RANK, NF-κB1 and TRAP proteins (1.11 ± 0.04, 1.29 ± 0.05, 1.38 ± 0.04, 1.24 ± 0.04, 1.13 ± 0.03, 1.34 ± 0.05, 1.12 ± 0.03, 1.24 ± 0.04, 1.61 ± 0.06) were increased (n P < 0.05); TRAF-6 protein expressions in F n 0.1 and Fn 1.6 groups (1.23 ± 0.04, 1.35 ± 0.03) were increased (n P < 0.05). When arsenic was used alone, compared with the control group (F n 0.0Asn 0.0), the expressions of RANK, TRAF-6, NF-κB1 proteins were increased in Asn 0.5 group (n P < 0.05), the expressions of RANK and NFATc1 proteins were reduced in As n 12.5 group (n P < 0.05). When fluorine was combined with arsenic, at the same dose of fluorine, RANK protein expression in F n 0.1Asn 0.5 group and TRAF-6 protein expression in Fn 0.1Asn 12.5, Fn 0.4Asn 0.5, Fn 0.4Asn 2.5 groups, NF-κB1 protein expression in Fn 0.1Asn 0.5 Fn 0.4Asn 2.5, Fn 0.4Asn 12.5 groups, NFATc1 protein expression in Fn 0.1Asn 0.5 and Fn 0.4Asn 0.5 groups, TRAP protein expression in Fn 0.1Asn 12.5 group were higher than the corresponding fluorine groups alone (Fn 0.1, Fn 0.4, n P < 0.05), but lower than the sum of fluorine and arsenic alone. At the same dose of arsenic, RANK protein expression in F n 0.1Asn 12.5 group, TRAF-6 protein expression in Fn 0.1Asn 12.5 and Fn 0.4Asn 2.5 groups, and NF-κB1 protein expression in Fn 0.1Asn 12.5, Fn 0.4Asn 2.5, Fn 0.4Asn 12.5, and Fn 1.6Asn 2.5 groups, TRAP protein expression in Fn 1.6Asn 2.5 and Fn 1.6Asn 12.5 groups were higher than the corresponding arsenic groups alone (Asn 2.5, Asn 12.5, n P < 0.05), but lower than the sum of fluorine and arsenic alone. Fluorine had a major effect on the expressions of RANK, TRAF-6, NF-κB1, NFATc1, and TRAP proteins ( n F=3.41, 341.73, 66.01, 56.49, 147.40, n P < 0.05); arsenic also had a main effect on all protein indicators ( n F=686.71, 174.96, 107.32, 235.80, 331.37, n P < 0.05); the combined effect of fluorine and arsenic had an interaction effect on each protein indicator ( n F=50.39, 234.94, 116.72, 67.77, 36.56, n P < 0.05).n Conclusions:In the co-culture system of MC3T3-E1 and RAW264.7 cells, fluorine can activate TRAF-6-mediated expression of NF-κB1 signaling pathway-related proteins, thereby promoting osteoclast differentiation; the effects of arsenic on the expression of related proteins are not completely consistent. The interaction of fluorine and arsenic exposure on TRAF-6-mediated expression of NF-κB1 signaling pathway-related proteins is mainly antagonistic.